Clinical and genetic diagnosis for 26 paitents with hereditary spherocytosis / 中南大学学报(医学版)

OBJECTIVES@#Hereditary spherocytosis (HS) is the most common hereditary defect of the red cell membrane, mainly characterized by anemia, jaundice, and splenomegaly. Due to the atypical clinical manifestations and negative family history of some patients, as well as the low sensitivity and specificity of traditional laboratory examinations, it is easy for it to escape diagnosis or be misdiagnosed. At present, it has been confirmed that the mutation of ANK1, SPTB, SPTA1, SLC4A1 and EPB42 genes can cause the deletion of their corresponding coding proteins, and thus lead to the defect of erythrocyte membrane. This study aims to analyze the feasibility and clinical application value of HS gene diagnosis.@*METHODS@#Data of 26 patients from Hunan, China with HS admitted to the Department of Hematology, Second Xiangya Hospital of Central South University from January 2018 to September 2021 were retrospectively collected, and their clinical manifestations and results of laboratory examinations were analyzed. Next-generation sequencing (NGS) combined with Sanger sequencing were applied. The mutation of HS pathogenic gene and the variation of uridine diphosphate-glucuronosyl transferase 1 family polypeptide A1 (UGT1A1), a key enzyme in the regulation of bilirubin metabolism, were detected. The results of pathogenic gene variations were interpreted pathogenic gene variations in accordance with the Standards and guidelines for the interpretation of sequence variants published by the American College of Medical Genetics and Genomics (ACMG). The clinical characteristics of patients with different gene variants were analyzed, and the clinical diagnosis and genetic diagnosis were compared.@*RESULTS@#Among the 26 patients with HS, there were 23 cases of anemia, 25 cases of jaundice, 24 cases of splenomegaly, and 14 cases of cholelithiasis. There were 16 cases with family history and 10 cases without family history. The results of HS mutation test were positive in 25 cases and negative in 1 case. A total of 18 heterozygous mutations of HS pathogenic genes were detected in 19 families, among which 14 were pathogenic, 1 was likely pathogenic and 3 were of unknown significance. SPTB mutations (12) and ANK1 mutations (4) were the most common. The main variation types were nonsense mutation (9). There were no significant differences in peripheral blood cell parameters and hemolysis indicators between the SPTB mutant group and the ANK1 mutant group (all P>0.05). The rate of splenectomy in ANK1 mutation group was higher than that in SPTB mutation group, and the difference was statistically significant (χ2=6.970, P=0.014). There were no significant differences in peripheral blood cell parameters and hemolysis indicators among different mutation types (nonsense mutation, frameshift mutation, splice site mutation and missense mutation) (all P>0.05). Among the 18 clinically confirmedpatients, there were 17 cases whose diagnosis is consistent with the genetic diagnosis. Eight patients were clinically suspected, and all of them were confirmed by detection of HS gene mutation. Twenty-four patients with HS underwent UGT1A1 mutation detection, among which 5 patients carried UGT1A1 mutation resulting in a decrease in enzyme activity, and 19 patients had normal enzyme activity. The level of total bilirubin (TBIL) in the group with reduced enzyme activity was higher than that in the group with normal enzyme activity, and the difference was statistically significant (U=22, P=0.038).@*CONCLUSIONS@#Most patients with HS have anemia, jaundice and splenomegaly, often accompanied by cholelithiasis. SPTB and ANK1 mutations are the most common mutations in HS pathogenic genes among patients in Hunan, China, and there was no significant correlation between genotype and clinical phenotype. Genetic diagnosis is highly consistent with clinical diagnosis. The decrease of UGT1A1 enzyme activity can lead to the aggravation of jaundice in HS patients. Clinical combined gene diagnosis is beneficial for the rapid and precision diagnosis of HS. The detection of UGT1A1 enzyme activity related gene variation plays an important role in evaluation of HS jaundice.

X-ray ionizing radiation up-regulates c-Myc expression and promotes epithelial-mesenchymal transition in lung cancer cell line A549 / 中国病理生理杂志

AIM:To investigate the effect of ionizing radiation on epithelial-mesenchymal transition in lung cancer cell line A549 and its possible mechanism.METHODS:The lung cancer A549 cells were irradiated with different doses (0 Gy, 1 Gy, 2 Gy, 4 Gy and 8 Gy) of X-ray for different time.The morphological changes of the cells were observed under inverted microscope at time points of 12 h, 24 h and 48 h.The expression of vimentin, N-cadherin, E-cadherin and transcription factor c-Myc was detected by Western blot at the time points of 12 h and 24 h.RESULTS:After ionizing radiation, the contours of the A549 cells were unclear, the protrusions increased, and the edges were irregular, with fried egg-like collapses.The mesenchymal morphology of the A549 cells was most obvious after irradiation at 8 Gy for 48 h.Compared with 0 Gy irradiation group, the expression of vimentin was down-regulated seemingly 12 h after irradiation, but up-regulated in 2 Gy, 4 Gy and 8 Gy irradiation groups for 24 h, and the most obvious effect was observed in 2 Gy irradiation group (P<0.01).Compared with 0 Gy irradiation group, the expression of N-cadherin was up-regulated in 1 Gy, 2 Gy and 4 Gy irradiation groups for 24 h (P<0.05), while the expression of E-cadherin was not influenced.The up-regulation of vimentin expression in lung cancer cell line A549 was positively correlated with c-Myc expression.CONCLUSION:Ionizing radiation may promotes epithelial-mesenchymal transition in the lung cancer cell line A549 by up-regulating the c-Myc expression.

Effect of goal-directed fluid therapy on the intestinal blocking of the elderly patients with colon cancer surgery / 实用医学杂志

Objective To evaluate the effects of two fluid therapy methods guided by SVV and CVP re-spectively on intestinal barrier of colon cancer surgery with elderly patients. Methods 50 elderly patients with ASA Ⅱ~Ⅲ level were randomly divided into Group S (fluid therapy guided by SVV) and Group C (convention-al fluid therapy). Each group has 25 cases. Patients in group S were treated under goal-directed fluid infusion strategy with target of 10%≤SVV≤12%. Group C received conventional infusion characterized by monitoring central venous pressure (CVP) and mean arterial pressure (MAP). Fluid volume, surgery time, urine volume, blood loss, exhausting time, post-operative hospitalization days in Group S and Group C were recorded. MAP, HR, CVP, SVV, HCO3- of patients were recorded at the same time before anesthesia (T0), beginning of surgery (T1), one hour after surgery (T2) and ending of surgery (T3) respectively. Venous blood samples were collected at the time points of T0, T1, T2 and T3 to detect DAO and D-lactic acid levels. Results MAP and CVP at time points of T2 and T3 in Group S were obviously lower than that in Group C; total intraoperative fluid volume in Group S was obviously less than that in Group C (P < 0. 05); postoperative exhausting time and hospitalization days in Group S decreased significantly (P < 0.05) and the content of DAO and D-lactic acid in Group S were lower than that in Group C (P < 0.05). Conclusion For elderly patients with colon cancer surgery, fluid thera-py guided by SVV is better than conventional fluid therapy guided by CVP in protecting intestinal barrier.